Evidence map›Paper›PMID 35755943›Full record

ArticleMolecular therapy. Methods & clinical development2022

PAM-flexible dual base editor-mediated random mutagenesis and self-activation strategies to improve CRISPRa potency.

Cia-Hin Lau, Siping Huang, Raymond H W Lam, Chung Tin

Open access · goldAbstract read
In one paragraph

Article in Molecular therapy. Methods & clinical development, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
0.3field-weighted citation impact, top 49% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed, 3 citations in OpenAlex.

  1. Review
  2. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors at 1 institution in 1 country.

Cia-Hin LauDepartment of Biomedical Engineering, City University of Hong Kong, Room P6416, Yeung Kin Man Academic Building, 83 Tat Chee Avenue, Kowloon Tong, Hong Kong, SAR, China.
Siping HuangDepartment of Biomedical Engineering, City University of Hong Kong, Room P6416, Yeung Kin Man Academic Building, 83 Tat Chee Avenue, Kowloon Tong, Hong Kong, SAR, China.
Raymond H W LamDepartment of Biomedical Engineering, City University of Hong Kong, Room P6416, Yeung Kin Man Academic Building, 83 Tat Chee Avenue, Kowloon Tong, Hong Kong, SAR, China.
Chung TinDepartment of Biomedical Engineering, City University of Hong Kong, Room P6416, Yeung Kin Man Academic Building, 83 Tat Chee Avenue, Kowloon Tong, Hong Kong, SAR, China.
City University of Hong Kong · HK

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

VP64 is the smallest transactivation domain that can be packaged together with the sgRNA into a single adeno-associated virus (AAV) vector. However, VP64-based CRISPRa often exerts modest activation to the target gene when only one sgRNA is used. Herein, we used PAM-flexible dual base editor-mediated mutagenesis and self-activation strategies to derive VP64 variants with gain-of-function mutations. First, we generated an HEK293FT transgenic clone to stably expressing pTK-CRISPRa-GFP. The sgRNA of CRISPRa was designed to target the TK promoter, thereby allowing self-activation of CRISPRa-GFP. Base editors were then used to randomly mutagenesis VP64 in this transgenic cell. VP64 with enhanced potency would translate into increment of GFP fluorescence intensity, thereby allowing positive selection of the desired VP64 mutants. This strategy has enabled us to identify several VP64 variants that are more potent than the wild-type VP64. ΔCRISPRa derived from these VP64 variants also efficiently activated the endogenous promoter of anti-aging and longevity genes (

Indexed as

adeno-associated virusbase editingdual base editornCas9-NGpositive feedbackSpRYVP16VP64

Identifiers

PMID35755943
PMCPMC9198377
OpenAlexW4281747000

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.